JAX Mouse Strain Datasheet - 014093

STOCK Tg(tetO-CHRM3*)1Blr/J

Stock No:: 014093 |; TRE-hM3Dq

Transgenic

Repository Live

Overview

Also Known As: TRE-hM3Dq

These TRE-hM3Dq transgenic mice may be bred to generate Tet-Off/Tet-On mutant mice with conditional (inducible/reversible) expression of hM3Dq; a mutant G protein-coupled receptor (GPCR) that activates the canonical G_q pathway specifically following administration of the pharmacologically inert molecule clozapine-N-oxide (CNO). TRE-hM3Dq transgenic mice may be useful for chemogenetic/pharmacogenetic applications to study receptor-specific functions or general downstream cellular signaling emanating from the activated GPCR.

Donating Investigator

Bryan L Roth, University of North Carolina at Chapel Hill

Genetic overview

Genetic Background	Generation
	?+pN2F1 (20-JUL-16)

Tg(tetO-CHRM3*)1Blr

Allele Type
Transgenic (Inducible, Inserted expressed sequence)

View Genetics

Research Applications

Cancer Research
Cell Biology Research
Diabetes and Obesity Research
Immunology, Inflammation and Autoimmunity Research
Neurobiology Research
Research Tools

View All Research Applications

Base Price

Starting at:

$246.50 Domestic price for female

$320.50 Domestic price for breeder pair

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Details

Detailed Description

Hemizygous TRE-hM3Dq transgenic mice are viable and fertile, with no reported phenotypic abnormalities. The TRE-hM3Dq transgene has a modified Tet response element (TRE or tetO) upstream of the mutant G protein-coupled receptor, hM3Dq (a human muscarinic 3 receptor with two amino acid substitutions (Y149C^3.33/A239G^5.46) that abolish receptor affinity for the native ligand, acetylcholine (ACh), but allow receptor binding and subsequent activation by the small pharmacologically inert molecule clozapine-N-oxide (CNO)). When bred with another mouse expressing tetracycline-controlled transactivator protein (tTA) or reverse tetracycline-controlled transactivator protein (rtTA), hM3Dq expression in the resulting double mutant offspring can be regulated with tetracycline or its analog doxycycline (dox). As designed, TRE-hM3Dq transgenic mice have no reported levels of hM3Dq activity in tTA(+dox) or rtTA(-dox) cell types. In TRE-hM3Dq transgenic tTA(-dox) or TRE-hM3Dq transgenic rtTA(+dox) cells types, hM3Dq expression results in activation of the canonical G_q pathway only following administration of CNO.

These TRE-hM3Dq transgenic mice may be useful to study receptor-specific functions or general downstream cellular signaling emanating from the activated G protein-coupled receptor (GPCR). For example, when bred to a strain expressing tTA in forebrain neurons (Camk2a-tTA; see Stock No. 003010), these transgenic mice are a model allowing in vivo chemical control of neuronal activity, neuronal firing, and non-neuronal signaling.

In addition, breeding TRE-hM3Dq mice with cfos-htTA mice (Stock No. 018306) generates double transgenic offspring with hM3Dq expression in excitatory neurons that are sufficiently active to drive the c-fos promoter. Those hM₃D_q^fos double transgenic mice allow CNO ligand-induced, artificial reactivation of neurons associated with a specific memory. Temporal modulation may also be employed using dox. Such hM₃D_q^fos double transgenic mice allow one to generate and artificially-activate a synthetic memory trace, and may be useful in studying the spatial pattern of activity at the time of learning and retrieval.

Of note, several chemogenetic/pharmacogenetic tool strains are available from The Jackson Laboratory Repository; including these Tet-responsive TRE-hM3Dq transgenic mice (Stock No. 014093), the adora2A-rM3Ds transgenic mice (Stock No. 017863), the Tet-responsive TRE-hM4Di transgenic mice (Stock No. 024114), the Cre-inducible R26-LSL-Gi-DREADD mice (R26-hM4Di/mCitrine; Stock No. 026219) and the Cre-inducible CAG-LSL-Gq-DREADD mice (Tg-hM3Dq/mCitrine [formerly R26-LSL-Gq-DREADD]; Stock No. 026220).

For CNO protocol, see the detailed protocol for dissolving CNO obtained from the attachments section of the Designer Receptors Exclusively Activated by Designer Drugs DREADD wiki webpage.

Development

The TRE-HA-hM3Dq transgene was designed with a modified Tet response element (TRE or (tetO); described in detail below), a hemagglutinin epitope tag (HA), the hM3Dq sequence, and an SV40 polyA signal. The hM3Dq sequence is a G_q-coupled human M3 muscarinic DREADD (designer receptor exclusively activated by designer drug). To create the hM3Dq sequence, the wildtype human muscarinic 3 receptor (CHRM3) sequence was modified via site-directed mutagenesis to harbor two amino acid substitutions (Y149C^3.33/A239G^5.46) that abolish receptor affinity for the native ligand, acetylcholine (ACh), but allow receptor binding and subsequent activation by the small drug-like molecule clozapine-N-oxide (CNO).
This 2.36 kb TRE-HA-hM3Dq transgene was injected into the pronuclei of B6SJL hybrid mouse oocytes. Founder TRE-hM3Dq mice (line 1) were bred with B6;CBA-Tg(Camk2a-tTA)1Mmay/J (see Stock No. 003010) to establish the colony. TRE-hM3Dq transgenic mice were bred together or to wildtype (noncarrier) mice from the colony for approximately eight generations, and then crossed with C57BL/6J mice for one or two generations. The Camk2a-tTA transgene was removed during these breedings. Upon arrival at The Jackson Laboratory Repository, TRE-hM3Dq transgenic mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.

Additional information on hM3Dq:
More information on hM3Dq or other DREADDs may be available at the University of North Carolina Designer Receptors Exclusively Activated by Designer Drugs DREADD wiki webpage.

Additional information on TRE promoter:
The TRE promoter used in these transgenic mice is the Tet-responsive P_tight promoter. This P_tight promoter contains a modified Tet response element (TRE_mod), which consists of seven direct repeats of a 36-bp sequence each containing the 19-bp tet operator sequence (tetO). The TRE_mod is just upstream of a minimal human cytomegalovirus (CMV) promoter (P_minCMVΔ), which lacks the enhancer that is part of the complete CMV promoter. Consequently, P_tight is silent in the absence of binding of TetR or rTetR to the tetO sequences.

Expression Data

Expressed Gene	CHRM3, cholinergic receptor muscarinic 3, human
Site of Expression

Control Suggestions

Noncarrier

Approximate Controls

100011 B6CBAF1/J

Additional Information

Considerations for Choosing Controls

Selected References

Alexander GM; Rogan SC; Abbas AI; Armbruster BN; Pei Y; Allen JA; Nonneman RJ; Hartmann J; Moy SS; Nicolelis MA; McNamara JO; Roth BL. 2009. Remote control of neuronal activity in transgenic mice expressing evolved G protein-coupled receptors. Neuron 63(1):27-39. PubMed: 19607790 MGI: J:154952

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Genetics

Tg(tetO-CHRM3*)1Blr

Allele Symbol: Tg(tetO-CHRM3*)1Blr

Allele Name	transgene insertion 1, Bryan Roth
Allele Type	Transgenic (Inducible, Inserted expressed sequence)
Allele Synonym(s)	TRE-HA-hM3Dq; TRE-hM3Dq; tetO-hM3Dq
Gene Symbol and Name	Tg(tetO-CHRM3*)1Blr, transgene insertion 1, Bryan Roth
Gene Synonym(s)
Promoter	CMV, cytomegalovirus, human
Promoter	tetO, tet operator,
Expressed Gene	CHRM3, cholinergic receptor muscarinic 3, human
Strain of Origin	(C57BL/6 x SJL)F1
Chromosome	UN
Molecular Note	The Tet-responsive P promoter which consists of seven direct repeats of 36-bp sequence each containing the 19-bp tet operator sequence (tetO) is just upstream of a minimal human cytomegalovirus (CMV) promoter. This is upstream of a hemagglutinin epitop tag and a Gq-coupled human M3 muscarinic receptor. The wildtype human muscarinic 3 receptor sequence was modified via site-directed mutagenesis to harbor two amino acid substitutions (Y149C and A239G) that abolish receptor affinity for the native ligand, acetylcholine (ACh), but allow receptor binding and subsequent activation by the small drug-like molecule clozapine-N-oxide (CNO).
Mutations Made By	Bryan Roth, University of North Carolina at Chapel Hill

Disease/Phenotype

Disease Terms

Potential model based on transgenic expression of an ortholog of a human gene that is associated with this disease. Phenotypic similarity to the human disease has not been tested.

Prune Belly Syndrome; PBS (CHRM3)

Research Areas By Genotype

This mouse can be used to support research in many areas including:

Cancer Research
- Growth Factors/Receptors/Cytokines
Cell Biology Research
- Signal Transduction
Diabetes and Obesity Research
- Insulin Receptors and Growth Factors
Immunology, Inflammation and Autoimmunity Research
- Growth Factors/Receptors/Cytokines
- Intracellular Signaling Molecules
Neurobiology Research
- Optogenetic and Chemogenetic tools
  - Chemogenetic tools
- Receptor Defects
  - G protein-coupled receptor
- Tet Expression System
  - tTA/rtTA Responsive Strains
Research Tools
- Cancer Research
  - Tetop Tet System
- Cardiovascular Research
  - Tetop Tet System
- Genetics Research
  - Mutagenesis and Transgenesis: Tetop Tet System
- Neurobiology Research
  - Tetop Tet System
- Reproductive Biology Research
  - Tetop Tet System
- Tet Expression Systems
  - tTA/rtTA Responsive Strains

References

Alexander GM; Rogan SC; Abbas AI; Armbruster BN; Pei Y; Allen JA; Nonneman RJ; Hartmann J; Moy SS; Nicolelis MA; McNamara JO; Roth BL. 2009. Remote control of neuronal activity in transgenic mice expressing evolved G protein-coupled receptors. Neuron 63(1):27-39. PubMed: 19607790 MGI: J:154952

Additional - Tg(tetO-CHRM3*)1Blr related

Garner AR; Rowland DC; Hwang SY; Baumgaertel K; Roth BL; Kentros C; Mayford M. 2012. Generation of a synthetic memory trace. Science 335(6075):1513-6. PubMed: 22442487 MGI: J:181832
McClain JL; Fried DE; Gulbransen BD. 2015. Agonist-evoked Ca2+ signaling in enteric glia drives neural programs that regulate intestinal motility in mice. Cell Mol Gastroenterol Hepatol 1(6):631-645. PubMed: 26693173 MGI: J:239994

Pricing & Availability

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Cryorecovery - Pricing

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Also Known As: TRE-hM3Dq

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Approximate Controls

Additional Information

Selected References

Tg(tetO-CHRM3*)1Blr

Allele Symbol: Tg(tetO-CHRM3*)1Blr

Disease Terms

Potential model based on transgenic expression of an ortholog of a human gene that is associated with this disease. Phenotypic similarity to the human disease has not been tested.

Research Areas By Genotype

This mouse can be used to support research in many areas including:

References

Additional - Tg(tetO-CHRM3*)1Blr related

Genotyping Protocols

Dietary Information

Breeding Considerations

Mating System

Animal Health Reports

Live Mouse

Breeder Pair

Cryorecovery - Pricing

Progeny testing is not required

Payment Terms and Conditions

The Jackson Laboratory's Genotype Promise

Terms of Use

Additional Use Restrictions Apply

Licensing Information

JAX® Mice, Products & Services Conditions of Use

No Warranty

Credit for PRODUCTS or SERVICES

Animal Care and Use for SERVICES

No Liability